Algal omega 7 compositions

ABSTRACT

Provided herein are exemplary algal omega 7 compositions, including algal fatty acid compositions comprising by dry weight from about approximately 0.5% to about approximately 99% C16:1 n7 palmitoleic acid (POA). Such algal compositions may also include (either individually or any combination of) by dry weight: from about approximately 0% to about approximately 20% saturated fatty acids; from about approximately 0% to about approximately 99% arachidonic acid; from about approximately 0% to about 99% docosahexaenoic acid; and/or from about approximately 0% to about approximately 99% eicosapentaenoic acid. Further exemplary algal fatty acid compositions may include by dry weight about approximately 90% POA, less than about approximately 20% saturated fatty acids, less than about approximately 10% ARA, substantially no DHA, and less than about approximately 10% EPA.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application claims the benefit and priority of U.S.Provisional Patent Application Ser. No. 61/800,114 filed on Mar. 15,2013 and titled “(EPA) Algal Biomass and Oil Compositions and Impact onHealth,” which is hereby incorporated by reference.

The present application claims the benefit and priority of U.S.Provisional Patent Application Ser. No. 61/800,029 filed on Mar. 15,2013 and titled “Microalga Species and Industrial Applications,” whichis hereby incorporated by reference.

The present application is related to U.S. Non-Provisional patentapplication Ser. No. ______, filed on ______ concurrently with thepresent application and titled “Algal Oil Compositions,” which is herebyincorporated by reference.

The present application is related to U.S. Non-Provisional patentapplication Ser. No. ______, filed on ______ concurrently with thepresent application and titled “Conversion of Free Fatty Acids to EthylEsters,” which is hereby incorporated by reference.

The present application is related to U.S. Non-Provisional patentapplication Ser. No. ______, filed on ______ concurrently with thepresent application and titled “Algal Omega 7 and Algal Omega 3 BlendCompositions,” which is hereby incorporated by reference.

The present application is related to U.S. Non-Provisional patentapplication Ser. No. ______, filed on ______ concurrently with thepresent application and titled “Compositions and Methods for Utilizationof Algal Compounds,” which is hereby incorporated by reference.

BACKGROUND OF THE INVENTION Field of the Invention

This invention relates to algal biochemistry, and more specifically, toalgal omega 7 compositions.

SUMMARY OF THE INVENTION

Provided herein are exemplary algal omega 7 compositions, includingalgal fatty acid compositions comprising by dry weight from aboutapproximately 0.5% to about approximately 99% C16:1 n7 palmitoleic acid(POA). Such algal compositions may also include (either individually orany combination of) by dry weight: from about approximately 0% to aboutapproximately 10% saturated fatty acids; from about approximately 0% toabout approximately 99% arachidonic acid; from about approximately 0% toabout 99% docosahexaenoic acid; and/or from about approximately 0% toabout approximately 99% eicosapentaenoic acid.

Provided herein are also exemplary algal omega 7 compositions, includingalgal fatty acid compositions comprising by dry weight from aboutapproximately 0.5% to about approximately 99% C16:1 n7 palmitoleic acid(POA). Such algal compositions may also include (either individually orany combination of) by dry weight: from about approximately 0% to aboutapproximately 10% saturated fatty acids; from about approximately 0% toabout approximately 10% arachidonic acid; substantially no (i.e. lessthan approximately 0.5%) docosahexaenoic acid; and/or from aboutapproximately 0% to about approximately 10% eicosapentaenoic acid.

Further exemplary algal fatty acid compositions may include by dryweight about approximately 90% palmitoleic acid, less than aboutapproximately 10% saturated fatty acids, less than about approximately10% arachidonic acid, substantially no docosahexaenoic acid, and lessthan about approximately 10% eicosapentaenoic acid.

The algal palmitoleic acid compositions described herein may bedesaturated (i.e. removing the saturated fatty acids from themonounsaturated and/or polyunsaturated fatty acids) from saturated algalcompositions comprising by total weight approximately 50% POA,approximately 50% palmitic acid (PA) and substantially no DHA. Thesaturated algal compositions may result from the processing of totalalgal oil compositions comprising by total weight approximately 30% EPA,approximately 27% POA, approximately 23% PA, less than approximately 10%ARA, and substantially no DHA.

The saturated algal compositions may also result from the processing oftotal algal oil compositions comprising by total weight approximately30% EPA, approximately 27% POA, less than approximately 20% saturatedfats, less than approximately 10% ARA, and substantially no DHA.

The saturated algal compositions may also result from the processing oftotal algal oil compositions comprising by total weight from betweenapproximately 0% EPA and 99% EPA, from between approximately 0% POA and99% POA, less than approximately 20% saturated fats, from betweenapproximately 0% ARA and 99% ARA, and from between approximately 0% DHAand 99% DHA.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows an actual fatty acid (in ethyl ester form) profile for anexemplary total algal oil composition.

FIG. 2 shows a flow chart for an exemplary method of unsaturating anexemplary saturated fatty acyl moieties (FAMs) rich algal composition.

FIG. 3 shows the actual content of 16:0 ethyl ester (palmitic acid)expressed as mg/g ethyl ester in the liquid phase at each of steps210-250 as described in connection with FIG. 2.

FIG. 4 shows an actual fatty acid profile for an exemplary fatty acidcomposition comprising mostly C16:1 n7 palmitoleic acid that resultedafter processing an exemplary total algal oil composition such as thatshown in FIG. 1 by the exemplary method shown and described inconnection with FIG. 2.

FIG. 5 is a data table for an actual fatty acid profile for an exemplarytotal algal oil composition (in ethyl ester form), such as shown in FIG.1, an exemplary saturated algal composition (in ethyl ester form), andan exemplary fatty acid composition (in ethyl ester form) comprisingmostly C16:1 n7 palmitoleic acid, as shown in FIG. 4.

FIG. 6 is an actual saturated fat profile for an exemplary whole biomass(“WB”) (in a form of a fatty acid methyl ester), an exemplary crudebiomass (“Crude”) (in a form of a fatty acid methyl ester), an exemplaryalgal oil composition (“TAO”) (in ethyl ester form), an exemplary 16chain fatty acid composition (in ethyl ester form), and an exemplaryC16:1 n7 palmitoleic acid (in ethyl ester form).

DETAILED DESCRIPTION OF THE INVENTION

A fatty acid is a carboxylic acid with a long aliphatic tail (chain),which is either saturated or unsaturated. Most naturally occurring fattyacids have a chain of an even number of carbon atoms, from 4 to 28.Saturated fatty acids have no double bonds between carbon atoms.Unsaturated fatty acids have one or more double bonds between carbonatoms. When counting from the terminal methyl carbon toward the carbonylcarbon on an unsaturated fatty acid, the first double bond signifies theomega double bond, such as observed in omega 3, omega 6, or omega 7fatty acids.

Palmitoleic acid (POA) is an omega-7 monounsaturated fatty acid with a16-carbon chain with one double bond, denoted as C16:1 n7. A beneficialfatty acid, it has been shown to suppress inflammation. Dietary sourcesof omega-7 are found in animal and plant sources, including seabuckthorn berries, macadamia nuts, cold water fish and dairy fat. Thesesources, however, are not concentrated and/or purified sources of POAand often contain a mixed fatty acid profile of saturated andpolyunsaturated fats.

Palmitic acid (PA) is a saturated fatty acid with a 16-carbon chain andno double bonds, denoted as C16:0. Consumption of saturated fats such aspalmitic acid is believed to increase the risk of developing diabetes,obesity, stroke and cardiovascular diseases.

Alpha linolenic acid (ALA) is an omega-3 polyunsaturated fatty acid(PUFA) with an 18-carbon chain and three cis double bonds. The firstdouble bond is located at the third carbon from the methyl end of thefatty acid chain, denoted as C18:3 n3.

Arachidonic acid (ARA) is an omega-6 PUFA with a 20-carbon chain andfour cis-double bonds; the first double bond is located at the sixthcarbon from the omega end. ARA is also denoted as C20:4 n6. Examples ofdietary sources of omega-6 PUFAs include refined vegetable oils, such ascorn and soy oil, seeds and nuts and the oils extracted from them.Consumption is therefore sufficient in the average diet.

Eicosapentaenoic acid (EPA) is an omega-3 fatty acid PUFA with theconnotation C20:5 n3. It is a carboxylic acid with a 20-carbon chain andfive cis double bonds; the first double bond is located at the thirdcarbon from the omega end.

Docosahexaenoic acid (DHA) is an omega-3 fatty acid PUFA. It is acarboxylic acid with a 22-carbon chain and six cis double bonds; thefirst double bond is located at the third carbon from the omega end. DHAis also denoted as C22:6 n3.

Provided herein are exemplary algal omega 7 compositions, includingalgal fatty acid compositions comprising by dry weight from aboutapproximately 0.5% to about approximately 99% C16:1 n7 palmitoleic acid(POA). Such algal compositions may also include (either individually orany combination of) by dry weight: from about approximately 0% to aboutapproximately 20% saturated fatty acids; from about approximately 0% toabout approximately 99% arachidonic acid; from about approximately 0% toabout 99% docosahexaenoic acid; and/or from about approximately 0% toabout approximately 99% eicosapentaenoic acid.

The various exemplary algal omega 7 compositions provided herein maycomprise by dry weight from about approximately 0.5% to aboutapproximately 99% palmitoleic acid. Such algal compositions may alsoinclude (either individually or any combination of) by dry weight: fromabout approximately 0% to about approximately 20% saturated fatty acids;from about approximately 0% to about approximately 10% arachidonic acid;substantially no (i.e. less than approximately 0.5%) docosahexaenoicacid; and/or from about approximately 0% to about approximately 10%eicosapentaenoic acid.

Additionally, the various exemplary algal omega 7 compositions providedherein may further be in ethyl ester form. Such ethyl esters are derivedby reacting free fatty acids with ethanol. Called esterification, theresulting ethyl ester allows for the fractional distillation(concentration) of the long chain fatty acids at lower temperatures.This step allows for the selective concentration of the fatty acids tolevels greater than found in nature.

The ethyl ester forms of the various exemplary algal omega 7compositions provided herein may be converted to a triglyceride form byperforming an enzymatic reaction with the ethyl ester form in thepresence of glycerol, heating under a vacuum, and filtering out theenzymes. Per some exemplary methods, immobilized lipase enzymes may bethose isolated from Candida antarctica and/or commercially supplied byNovozyme or Sigma Aldrich.

The algal palmitoleic acid compositions described herein may bedesaturated (i.e. removing the saturated fatty acids from themonounsaturated and/or polyunsaturated fatty acids) from saturated algalcompositions comprising by total weight approximately 50% POA,approximately 50% palmitic acid (PA) and substantially no DHA. Thesaturated algal compositions may result from the processing of totalalgal oil compositions comprising by total weight approximately 30% EPA,approximately 27% POA, approximately 0% to 20% saturated fats, less thanapproximately 10% ARA, and substantially no DHA.

The saturated algal compositions may also result from the processing oftotal algal oil compositions comprising by total weight approximately30% EPA, approximately 27% POA, less than approximately 20% saturatedfats, less than approximately 10% ARA, and substantially no DHA.

The saturated algal compositions may also result from the processing oftotal algal oil compositions comprising by total weight from betweenapproximately 0% EPA and 99% EPA, from between approximately 0% POA and99% POA, less than approximately 10% saturated fats, from betweenapproximately 0% ARA and 99% ARA, and from between approximately 0% DHAand 99% DHA.

In various embodiments, various algae species may be the source of thecompositions provided herein. Algae are mostly aquatic photosyntheticorganisms that range from microscopic flagellate to giant kelp. Algaemay be loosely grouped into seven categories: Euglenophyta (euglenoids),Chrysophyta (golden-brown algae), Pyrrophyta (fire algae),Dinoflagellata, Chlorophyta (green algae), Rhodophyta (red algae),Paeophyta (brown algae), and Xanthophyta (yellow-green algae). Lipidextracted from any algae genus may be used in the various embodiments ofthe present invention, including Amphora, Anabaena, Anikstrodesmis,Botryococcus, Chaetoceros, Chlorella, Chlorococcum, Cyclotella,Cylindrotheca, Dunaliella, Emiliania, Euglena, Glossomastix,Haematococcus, Isochrysis, Monochrysis, Monoraphidium, Nannochloris,Nannochloropsis, Navicula, Nephrochloris, Nephroselmis, Nitzschia,Nodularia, Nostoc, Oochromonas, Oocystis, Oscillatoria, Pavlova,Phaeodactylum, Picochloris, Platymonas, Pleurochrysis, Porphyra,Pseudoanabaena, Pyramimonas, Scenedesmus, Stichococcus, Synechococcus,Synechocystis, Tetraselmis, Thalassiosira, and Trichodesmium.

FIG. 1 shows an actual fatty acid (in ethyl ester form) profile for anexemplary total algal oil composition that was produced and analyzed bythe present inventors. As illustrated by FIG. 1, exemplary total algaloil compositions may comprise by total weight approximately 30% EPA,approximately 27% palmitoleic acid, approximately approximately 0% to20% saturated fats, less than approximately 10% ARA, and substantiallyno DHA.

An exemplary total algal oil composition, such as the exemplary totalalgal oil composition shown in FIG. 1, may be processed via high vacuumdistillation, resulting in an exemplary algal composition comprisingsaturated fatty acyl moieties (FAMs) including by total weightapproximately 50% POA, approximately 50% PA and substantially no DHA.

FIG. 2 shows a flow chart for an exemplary method of unsaturating anexemplary saturated fatty acyl moieties (FAMs) rich algal composition.

At step 210, 100 milliliters (mls) of the exemplary saturated algalcomposition is chilled at 4° C. for 24 hours, resulting in formation oftransparent crystals as stoutly needles.

At step 220, the mixture comprising the transparent crystals isvacuum-filtered through a whatman #1 paper filter, and the cake isblotted with kimwipes to remove interstitial liquid.

At step 230, the filtered liquid phase is brought to −3° C. for 24hours, and a second crop of crystals is formed as described above.

At step 240, the filtered liquid phase is brought to −10° C. for 24hours, and a third crop of crystals is formed, these crystals beingsmaller and rounder than earlier crops.

At step 250, the filtered liquid phase is then brought to −20° C. for 24hours, and forms a turbid suspension which is filtered.

FIG. 3 shows the actual content of 16:0 ethyl ester (palmitic acid)expressed as mg/g (in ethyl ester form) in the liquid phase at each ofsteps 210-250 as described in connection with FIG. 2. Referring to FIG.3, each diamond represents the actual content of 16:0 ethyl ester(palmitic acid) expressed as mg/g (in ethyl ester form) in the liquidphase, beginning with step 210 for the diamond at the left, andprogressing to step 250 for the diamond at the right.

FIG. 4 shows an actual fatty acid profile for an exemplary fatty acidcomposition comprising mostly C16:1 n7 palmitoleic acid that resultedafter the present inventors processed an exemplary total algal oilcomposition such as that shown in FIG. 1 by the exemplary method shownand described in connection with FIG. 2.

As shown in FIG. 4, exemplary algal omega 7 compositions may compriseapproximately 90% palmitoleic acid, less than approximately 10%saturated fatty acids, less than approximately 2% arachidonic acid,substantially no docosahexaenoic acid, and less than approximately 10%eicosapentaenoic acid.

FIG. 5 is a data table for an actual fatty acid profile determined bythe present inventors for an exemplary total algal oil composition (inethyl ester form), such as shown in FIG. 1, an exemplary saturatedFAM-rich algal composition (in ethyl ester form), and an exemplary fattyacid composition (in ethyl ester form) comprising mostly C16:1 n7palmitoleic acid, as shown in FIG. 4.

The data table in FIG. 5 shows fatty acid type by milligrams per gram(mg/g) as found in the ethyl ester forms of the various algalcompositions. Remarkably, as shown in FIG. 5, the exemplary fatty acidcomposition (in ethyl ester form) comprising mostly C16:1 n7 palmitoleicacid, also comprises only 3.46% saturated fatty acids.

FIG. 6 is an actual saturated fat profile for an exemplary whole biomass(“WB”) (in a form of a fatty acid methyl ester), an exemplary crudebiomass (“Crude”) (in a form of a fatty acid methyl ester), an exemplarytotal algal oil composition (“TAO”) (in ethyl ester form), an exemplary16 chain fatty acid composition (in ethyl ester form), and an exemplaryC16:1 n7 palmitoleic acid (in ethyl ester form). The data is reflectedas % saturated fats to total saturated and non-saturated fats.

As shown in FIG. 6, the exemplary algal composition is less saturatedthan both the exemplary crude and whole algal biomass compositions.

While various embodiments have been described above, it should beunderstood that they have been presented by way of example only, and notlimitation. Thus, the breadth and scope of a preferred embodiment shouldnot be limited by any of the above-described exemplary embodiments.

What is claimed is:
 1. An algal fatty acid composition comprising by dryweight approximately 0.5% to approximately 99% C16:1 n7 palmitoleic acidand less than approximately 20% saturated fatty acids.
 2. The algalfatty acid composition of claim 1, further comprising by dry weight lessthan approximately 10% arachidonic acid.
 3. The algal fatty acidcomposition of claim 1, further comprising by dry weight substantiallyno docosahexaenoic acid.
 4. The algal fatty acid composition of claim 1,further comprising by dry weight less than approximately 10%eicosapentaenoic acid.
 5. The algal fatty acid composition of claim 1,as unsaturated from a saturated fatty acyl moiety-rich algal compositioncomprising by total weight approximately 50% POA, approximately 50% PAand substantially no DHA.
 6. The saturated fatty acyl moiety-rich algalcomposition of claim 5, as processed from a total algal oil compositioncomprising by total weight approximately 30% EPA, approximately 27% POA,approximately 0% to 20% saturated fats, less than approximately 10% ARA,and substantially no DHA.
 7. An algal fatty acid composition comprisingby dry weight greater than approximately 70% palmitoleic acid and lessthan approximately 20% saturated fatty acids.
 8. The algal fatty acidcomposition of claim 7, further comprising by dry weight less thanapproximately 10% arachidonic acid.
 9. The algal fatty acid compositionof claim 7, further comprising by dry weight substantially nodocosahexaenoic acid.
 10. The algal fatty acid composition of claim 7,further comprising by dry weight less than approximately 10%eicosapentaenoic acid.
 11. The algal fatty acid composition of claim 7,as processed from a saturated fatty acyl moiety-rich algal compositioncomprising by total weight approximately 50% POA, approximately 50% PAand substantially no DHA.
 12. The saturated fatty acyl moiety-rich algalcomposition of claim 11, as processed from a total algal oil compositioncomprising by total weight approximately 30% EPA, approximately 27% POA,approximately 0% to 20% saturated fats, less than approximately 10% ARA,and substantially no DHA.
 13. An algal fatty acid composition comprisingby dry weight greater than approximately 80% palmitoleic acid and lessthan approximately 20% saturated fatty acids.
 14. The algal fatty acidcomposition of claim 13, further comprising by dry weight less thanapproximately 10% arachidonic acid.
 15. The algal fatty acid compositionof claim 13, further comprising by dry weight substantially nodocosahexaenoic acid.
 16. The algal fatty acid composition of claim 13,further comprising by dry weight less than approximately 10%eicosapentaenoic acid.
 17. The algal fatty acid composition of claim 13,as processed from a saturated fatty acyl moiety-rich algal compositioncomprising by total weight approximately 50% POA, approximately 50% PAand substantially no DHA.
 18. The saturated fatty acyl moiety-rich algalcomposition of claim 17, as processed from a total algal oil compositioncomprising by total weight approximately 30% EPA, approximately 27% POA,approximately 0% to 20% saturated fats, less than approximately 10% ARA,and substantially no DHA.
 19. An algal fatty acid composition comprisingby dry weight approximately 90% palmitoleic acid, less thanapproximately 20% saturated fatty acids, less than approximately 10%arachidonic acid, substantially no docosahexaenoic acid, and less thanapproximately 10% eicosapentaenoic acid.
 20. The algal fatty acidcomposition of claim 19, as processed from a saturated fatty acylmoiety-rich algal composition comprising by total weight approximately50% POA, approximately 50% PA and substantially no DHA.
 21. Thesaturated fatty acyl moiety-rich algal composition of claim 20, asprocessed from a total algal oil composition comprising by total weightapproximately 30% EPA, approximately 27% POA, approximately 0% to 20%saturated fats, less than approximately 10% ARA, and substantially noDHA.
 22. The algal fatty acid composition of claim 1, wherein thecomposition is in a form of an ethyl ester (EE), a mono, di, ortriacylglycerol (MAG, DAG, TAG), a phospholipid (PL), a galactolipid(GL), free fatty acid (FFA), or a sulfoquinovosyl diacylglycerol (SQDG).23. The algal fatty acid composition of claim 7, wherein the compositionis in a form of an ethyl ester (EE), a mono, di, or triacylglycerol(MAG, DAG, TAG), a phospholipid (PL), a galactolipid (GL), free fattyacid (FFA), or a sulfoquinovosyl diacylglycerol (SQDG).
 24. The algalfatty acid composition of claim 13, wherein the composition is in a formof an ethyl ester (EE), a mono, di, or triacylglycerol (MAG, DAG, TAG),a phospholipid (PL), a galactolipid (GL), free fatty acid (FFA), or asulfoquinovosyl diacylglycerol (SQDG).
 25. The algal fatty acidcomposition of claim 19, wherein the composition is in a form of anethyl ester (EE), a mono, di, or triacylglycerol (MAG, DAG, TAG), aphospholipid (PL), a galactolipid (GL), free fatty acid (FFA), or asulfoquinovosyl diacylglycerol (SQDG).